Confocal microscope apparatus

Abstract

A confocal microscope apparatus comprises a first optical scanning system which obtains a scan image of a sample using a laser beam from a first laser light source, a second optical scanning system which scans specific regions of a sample with a laser beam from a second laser light source that is different from the first laser light source, thereby causing a particular phenomenon, and a beam diameter varying mechanism which can change the beam diameter of the laser beam of at least one of the first optical scanning system and the second optical scanning system. With this configuration, the apparatus further comprises an excitation light intensity distribution calculator which calculates and stores the excitation light intensity distribution along a depth direction on the sample surface from the beam diameter of the laser beam output from the beam diameter varying mechanism.

What is claimed is: 
     
       1. A microscope apparatus comprising:
 a first optical system which illuminates a sample via an objective lens with incoherent light output from an incoherent light source, and detects fluorescence emitted from the sample via the objective lens; and 
 a second optical system which irradiates specific regions of the sample with a laser beam output from a laser light source, thereby causing a particular phenomenon; 
 wherein the first optical system comprises a Koehler illumination optical system; 
 wherein the second optical system comprises an optical scanning mechanism; and 
 wherein a depth position of a focal plane of the second optical system is substantially the same as a depth position of a focal plane of the first optical system. 
 
     
     
       2. The microscope apparatus according to  claim 1 , wherein the second optical system comprises a beam diameter varying mechanism which changes a beam diameter of the laser beam of the laser light source. 
     
     
       3. The microscope apparatus according to  claim 2 , further comprising an excitation light intensity distribution calculator which calculates and stores an excitation light intensity distribution along a depth direction on a surface of the sample from the beam diameter of the laser beam output from the beam diameter varying mechanism. 
     
     
       4. An observation method using a microscope, the method comprising:
 irradiating an excitation light to a sample via an optical system including a Koehler illumination optical system to acquire a fluorescent image of the sample; and 
 irradiating a laser light to cause a particular phenomenon at a desired position; 
 wherein a depth position of a focal plane of the irradiated excitation light is substantially the same as a depth position of a focal plane of the irradiated laser light. 
 
     
     
       5. The observation method according to  claim 4 , wherein the irradiating the laser light comprises adjusting an intensity distribution of the laser light along a depth direction on a surface of the sample by changing a beam diameter of the laser light.